PMID- 8641833
OWN - NLM
STAT- MEDLINE
DCOM- 19960716
LR  - 20181130
IS  - 0146-0404 (Print)
IS  - 0146-0404 (Linking)
VI  - 37
IP  - 7
DP  - 1996 Jun
TI  - Expression and release of tumor necrosis factor-alpha by explants of mouse 
      cornea.
PG  - 1302-10
AB  - PURPOSE: To elucidate a possible target of immunosuppressive agents widely used 
      in the treatment of corneal disorders, the authors determined whether corneal 
      cells are capable of expressing and releasing tumor necrosis factor-alpha (TNF 
      alpha) on lipopolysaccharide (LPS) stimulation, and they investigated whether TNF 
      alpha production can be modulated by pharmacologic agents. METHODS: Trephined 
      central corneas from C57BL/6 mice were kept in culture for 3 days. Release of TNF 
      alpha after a 24-hour stimulation with LPS (1 microgram/ml) into the culture 
      medium was determined both by bioassay and by enzyme-linked immunosorbent assay. 
      Expression of TNF alpha mRNA after 6-hour stimulation was examined by polymerase 
      chain reaction. Immunofluorescent staining on cryostat sections of cultured 
      corneas was performed to localize TNF alpha in the tissue. Corneal explants were 
      pretreated with immunosuppressive agents (prednisolone, budesonide, cyclosporin 
      A) for 48 hours, followed by 6-or 24-hour stimulation with LPS in the continuous 
      presence of the agents. RESULTS: Lipopolysaccharide stimulated TNF alpha release 
      into the culture medium. The addition of budesonide (10(-7) M) or prednisolone 
      (10(-6) M) significantly inhibited LPS-induced TNF alpha release, whereas 
      cyclosporin A (10(-7) - 10(-5) M) had no marked effect. Levels of TNF alpha mRNA 
      in corneal explants increased fivefold after stimulation with LPS. 
      Immunohistochemical staining revealed that TNF alpha was expressed in the 
      epithelial cells. Budesonide markedly decreased mRNA expression and abolished 
      immunostaining of TNF alpha stimulated by LPS. CONCLUSIONS: TNF alpha is produced 
      and released by the epithelial cells of mouse central cornea in response to LPS. 
      Contrary to cyclosporin A, corticosteroids such as prednisolone and budesonide 
      potently inhibit TNF alpha production.
FAU - Sekine-Okano, M
AU  - Sekine-Okano M
AD  - Department of Ophthalmology, University Hospital, Geneva, Switzerland.
FAU - Lucas, R
AU  - Lucas R
FAU - Rungger, D
AU  - Rungger D
FAU - De Kesel, T
AU  - De Kesel T
FAU - Grau, G E
AU  - Grau GE
FAU - Leuenberger, P M
AU  - Leuenberger PM
FAU - Rungger-Brandle, E
AU  - Rungger-Brandle E
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - 0 (Anti-Inflammatory Agents)
RN  - 0 (Culture Media)
RN  - 0 (DNA Primers)
RN  - 0 (Immunosuppressive Agents)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (RNA, Messenger)
RN  - 0 (Steroids)
RN  - 0 (Tumor Necrosis Factor-alpha)
SB  - IM
MH  - Animals
MH  - Anti-Inflammatory Agents/pharmacology
MH  - Base Sequence
MH  - Cornea/drug effects/*metabolism
MH  - Culture Media
MH  - DNA Primers/chemistry
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Epithelium/drug effects/metabolism
MH  - Escherichia coli
MH  - Female
MH  - Fluorescent Antibody Technique, Indirect
MH  - Immunosuppressive Agents/pharmacology
MH  - Lipopolysaccharides/antagonists & inhibitors/pharmacology
MH  - Macrophages, Peritoneal/metabolism
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Molecular Sequence Data
MH  - Organ Culture Techniques
MH  - Polymerase Chain Reaction
MH  - RNA, Messenger/biosynthesis
MH  - Steroids
MH  - Tumor Necrosis Factor-alpha/*biosynthesis/*metabolism
EDAT- 1996/06/01 00:00
MHDA- 1996/06/01 00:01
CRDT- 1996/06/01 00:00
PHST- 1996/06/01 00:00 [pubmed]
PHST- 1996/06/01 00:01 [medline]
PHST- 1996/06/01 00:00 [entrez]
PST - ppublish
SO  - Invest Ophthalmol Vis Sci. 1996 Jun;37(7):1302-10.